The construction and expression of a novel chimeric anti-DR5 antibody

Area of Science:

• Immunology
• Oncology
• Biotechnology

Background:

• Human tumor necrosis factor related apoptosis inducing ligand (TRAIL) shows promise for cancer therapy but can harm normal liver cells.
• Agonistic monoclonal antibodies (MAbs) against DR5 are being developed to selectively induce apoptosis in cancer cells and avoid TRAIL's side effects.
• Mouse-derived MAbs, like the previously developed mDRA6, can elicit harmful human anti-mouse antibody (HAMA) responses, limiting their clinical use.

Purpose of the Study:

• To engineer a chimeric anti-human DR5 MAb (cmDRA6) to overcome the limitations of mouse antibodies for potential cancer therapy.
• To generate and express cmDRA6 using variable region gene cloning and chimeric antibody expression vectors.

Main Methods:

• Variable region genes of mDRA6 were cloned into pCMV-VH and pCMV-VL expression vectors.
• Chimeric anti-human DR5 MAb (cmDRA6) expression vectors were created and introduced into 293T cells using Lipofectamine 2000.
• Expression levels of cmDRA6 were assessed using ELISA, and its antigen-binding capability was confirmed via ELISA and Western blot.

Main Results:

• Chimeric anti-human DR5 MAb (cmDRA6) was successfully expressed in 293T cells.
• ELISA and Western blot analyses confirmed that cmDRA6 binds to its target antigen, DR5.
• The study established a foundation for future research and clinical applications of cmDRA6.

Conclusions:

• The development of cmDRA6 represents a significant step towards creating safer and more effective antibody-based cancer therapeutics.
• Chimeric antibodies like cmDRA6 can mitigate HAMA responses, improving the therapeutic potential of DR5-targeting antibodies.